Baccosides content

REAGENTS & INSTRUMENT REQUIRED:

1. High Performance Liquid Chromatography
2. Water HPLC grade
3. Acetronitrile HPLC grade
4. Orthophosphoric Acid
5. Methanol LR grade
6. Baccosides working Standard

PROCEDURE:

Standard Preparation :

Weigh accurately about 250mg of in working standard into a 25mL volumetric flask. Add 20mL methanol and sonicate for 15 minutes. Make up to volume with methanol and Filter through 0.45μm filter paper.

Sample Preparation :

Weigh accurately about 250mg of sample into a 25mL volumetric flask . Add about 20mL solvents. Ultrasonicate for 15 minutes cool and make up the volume with methanol. Ensure that the concentration of active ingredient in sample solution is approximately similar to that of standard solution and filter through 0.45μm filter paper.

For Rawmaterial:  Take 10 gm of Herb powder (20#) and reflux in methanol for 1hrs twice.

Chromatographic Condition:

Mobile Phase               - 0.1% Orthophosphoric acid : Acetronitrile  (65 : 35) filter through 0.45μ and degas.

Column                        - Phenomenex luna 5µ, C18 (250mm X 4.6mm)

Mode                           – Isocratic

Injection Volume         – 20 µL

Flow rate                     – 1.0/1.5 mL /minute

Detector wavelength   – 205 nm

Condition                     - RT

Calculation:

                                    Area of sample   X   Concentration of std    X   purity of std

  % of  Baccosides =------------------------------------------------------------------------------------

                                       Area of standard    X   Concentration of sample